Veterinary Biosciences - Research Publications

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    Extracellular vesicles from early stage Plasmodium falciparum-infected red blood cells contain PfEMP1 and induce transcriptional changes in human monocytes
    Sampaio, NG ; Emery, SJ ; Garnham, AL ; Tan, QY ; Sisquella, X ; Pimentel, MA ; Jex, AR ; Regev-Rudzki, N ; Schofield, L ; Eriksson, EM (WILEY-HINDAWI, 2018-05)
    Pathogens can release extracellular vesicles (EVs) for cell-cell communication and host modulation. EVs from Plasmodium falciparum, the deadliest malaria parasite species, can transfer drug resistance genes between parasites. EVs from late-stage parasite-infected RBC (iRBC-EVs) are immunostimulatory and affect endothelial cell permeability, but little is known about EVs from early stage iRBC. We detected the parasite virulence factor PfEMP1, which is responsible for iRBC adherence and a major contributor to disease severity, in EVs, only up to 12-hr post-RBC invasion. Furthermore, using PfEMP1 transport knockout parasites, we determined that EVs originated from inside the iRBC rather than the iRBC surface. Proteomic analysis detected 101 parasite and 178 human proteins in iRBC-EVs. Primary human monocytes stimulated with iRBC-EVs released low levels of inflammatory cytokines and showed transcriptomic changes. Stimulation with iRBC-EVs from PfEMP1 knockout parasites induced more gene expression changes and affected pathways involved in defence response, stress response, and response to cytokines, suggesting a novel function of PfEMP1 when present in EVs. We show for the first time the presence of PfEMP1 in early stage P. falciparum iRBC-EVs and the effects of these EVs on primary human monocytes, uncovering a new mechanism of potential parasite pathogenesis and host interaction.
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    New species of Cloacina von Linstow, 1898 (Nematoda: Strongyloidea) parasitic in the stomachs of wallaroos, Osphranter spp. (Marsupialia: Macropodidae) from northern Australia
    Beveridge, I ; Jex, A ; Tan, N ; Jabbar, A (SPRINGER, 2018-07)
    Three new species of the parasitic nematode genus Cloacina von Linstow, 1898 (Strongyloidea: Cloacininae) are described from the stomachs of wallaroos, Osphranter spp. (Marsupialia: Macropodidae), from northern Australia. Cloacina spearei n. sp. is described from O. robustus woodwardi (Thomas) and O. antilopinus (Gould) and is distinguished from congeners by the shape of the cephalic papillae, the shallow buccal capsule, the presence of an oesophageal denticle and the convoluted but non-recurrent vagina in the female. Cloacina longibursata n. sp. also from O. robustus woodwardi and O. antilopinus is distinguished from congeners by the elongate dorsal lobe of the bursa, with the origin of the lateral branchlets posterior to the principal bifurcation, in the features of the spicule tip, the lack of bosses lining the oesophagus and the absence of an oesophageal denticle. Cloacina crassicaudata n. sp., from the same two host species was formerly identified as C. cornuta (Davey & Wood, 1938). Differences in the cephalic cuticle (inflation lacking in the new species), the shape of the cephalic papillae, the dorsal oesophageal tooth and the spicule tips, as well as differences in the sequences of the internal transcribed spacers of the nuclear ribosomal DNA, indicate that this is an independent species. The geographical distribution of this species is disjunct with populations in both the Northern Territory and Queensland. Possible reasons for the disjunct distribution are discussed.
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    A communal catalogue reveals Earth's multiscale microbial diversity
    Thompson, LR ; Sanders, JG ; McDonald, D ; Amir, A ; Ladau, J ; Locey, KJ ; Prill, RJ ; Tripathi, A ; Gibbons, SM ; Ackermann, G ; Navas-Molina, JA ; Janssen, S ; Kopylova, E ; Vazquez-Baeza, Y ; Gonzalez, A ; Morton, JT ; Mirarab, S ; Xu, ZZ ; Jiang, L ; Haroon, MF ; Kanbar, J ; Zhu, Q ; Song, SJ ; Kosciolek, T ; Bokulich, NA ; Lefler, J ; Brislawn, CJ ; Humphrey, G ; Owens, SM ; Hampton-Marcell, J ; Berg-Lyons, D ; McKenzie, V ; Fierer, N ; Fuhrman, JA ; Clauset, A ; Stevens, RL ; Shade, A ; Pollard, KS ; Goodwin, KD ; Jansson, JK ; Gilbert, JA ; Knight, R ; Rivera, JLA ; Al-Moosawi, L ; Alverdy, J ; Amato, KR ; Andras, J ; Angenent, LT ; Antonopoulos, DA ; Apprill, A ; Armitage, D ; Ballantine, K ; Barta, J ; Baum, JK ; Berry, A ; Bhatnagar, A ; Bhatnagar, M ; Biddle, JF ; Bittner, L ; Boldgiv, B ; Bottos, E ; Boyer, DM ; Braun, J ; Brazelton, W ; Brearley, FQ ; Campbell, AH ; Caporaso, JG ; Cardona, C ; Carroll, J ; Cary, SC ; Casper, BB ; Charles, TC ; Chu, H ; Claar, DC ; Clark, RG ; Clayton, JB ; Clemente, JC ; Cochran, A ; Coleman, ML ; Collins, G ; Colwell, RR ; Contreras, M ; Crary, BB ; Creer, S ; Cristol, DA ; Crump, BC ; Cui, D ; Daly, SE ; Davalos, L ; Dawson, RD ; Defazio, J ; Delsuc, F ; Dionisi, HM ; Dominguez-Bello, MG ; Dowell, R ; Dubinsky, EA ; Dunn, PO ; Ercolini, D ; Espinoza, RE ; Ezenwa, V ; Fenner, N ; Findlay, HS ; Fleming, ID ; Fogliano, V ; Forsman, A ; Freeman, C ; Friedman, ES ; Galindo, G ; Garcia, L ; Alexandra Garcia-Amado, M ; Garshelis, D ; Gasser, RB ; Gerdts, G ; Gibson, MK ; Gifford, I ; Gill, RT ; Giray, T ; Gittel, A ; Golyshin, P ; Gong, D ; Grossart, H-P ; Guyton, K ; Haig, S-J ; Hale, V ; Hall, RS ; Hallam, SJ ; Handley, KM ; Hasan, NA ; Haydon, SR ; Hickman, JE ; Hidalgo, G ; Hofmockel, KS ; Hooker, J ; Hulth, S ; Hultman, J ; Hyde, E ; Ibanez-Alamo, JD ; Jastrow, JD ; Jex, AR ; Johnson, LS ; Johnston, ER ; Joseph, S ; Jurburg, SD ; Jurelevicius, D ; Karlsson, A ; Karlsson, R ; Kauppinen, S ; Kellogg, CTE ; Kennedy, SJ ; Kerkhof, LJ ; King, GM ; Kling, GW ; Koehler, AV ; Krezalek, M ; Kueneman, J ; Lamendella, R ; Landon, EM ; Lane-deGraaf, K ; LaRoche, J ; Larsen, P ; Laverock, B ; Lax, S ; Lentino, M ; Levin, II ; Liancourt, P ; Liang, W ; Linz, AM ; Lipson, DA ; Liu, Y ; Lladser, ME ; Lozada, M ; Spirito, CM ; MacCormack, WP ; MacRae-Crerar, A ; Magris, M ; Martin-Platero, AM ; Martin-Vivaldi, M ; Margarita Martinez, L ; Martinez-Bueno, M ; Marzinelli, EM ; Mason, OU ; Mayer, GD ; McDevitt-Irwin, JM ; McDonald, JE ; McGuire, KL ; McMahon, KD ; McMinds, R ; Medina, M ; Mendelson, JR ; Metcalf, JL ; Meyer, F ; Michelangeli, F ; Miller, K ; Mills, DA ; Minich, J ; Mocali, S ; Moitinho-Silva, L ; Moore, A ; Morgan-Kiss, RM ; Munroe, P ; Myrold, D ; Neufeld, JD ; Ni, Y ; Nicol, GW ; Nielsen, S ; Nissimov, JI ; Niu, K ; Nolan, MJ ; Noyce, K ; O'Brien, SL ; Okamoto, N ; Orlando, L ; Castellano, YO ; Osuolale, O ; Oswald, W ; Parnell, J ; Peralta-Sanchez, JM ; Petraitis, P ; Pfister, C ; Pilon-Smits, E ; Piombino, P ; Pointing, SB ; Pollock, FJ ; Potter, C ; Prithiviraj, B ; Quince, C ; Rani, A ; Ranjan, R ; Rao, S ; Rees, AP ; Richardson, M ; Riebesell, U ; Robinson, C ; Rockne, KJ ; Rodriguezl, SM ; Rohwer, F ; Roundstone, W ; Safran, RJ ; Sangwan, N ; Sanz, V ; Schrenk, M ; Schrenzel, MD ; Scott, NM ; Seger, RL ; Seguin-Orlando, A ; Seldin, L ; Seyler, LM ; Shakhsheer, B ; Sheets, GM ; Shen, C ; Shi, Y ; Shin, H ; Shogan, BD ; Shutler, D ; Siegel, J ; Simmons, S ; Sjoling, S ; Smith, DP ; Soler, JJ ; Sperling, M ; Steinberg, PD ; Stephens, B ; Stevens, MA ; Taghavi, S ; Tai, V ; Tait, K ; Tan, CL ; Tas, N ; Taylor, DL ; Thomas, T ; Timling, I ; Turner, BL ; Urich, T ; Ursell, LK ; van der Lelie, D ; Van Treuren, W ; van Zwieten, L ; Vargas-Robles, D ; Thurber, RV ; Vitaglione, P ; Walker, DA ; Walters, WA ; Wang, S ; Wang, T ; Weaver, T ; Webster, NS ; Wehrle, B ; Weisenhorn, P ; Weiss, S ; Werner, JJ ; West, K ; Whitehead, A ; Whitehead, SR ; Whittingham, LA ; Willerslev, E ; Williams, AE ; Wood, SA ; Woodhams, DC ; Yang, Y ; Zaneveld, J ; Zarraonaindia, I ; Zhang, Q ; Zhao, H (NATURE PORTFOLIO, 2017-11-23)
    Our growing awareness of the microbial world's importance and diversity contrasts starkly with our limited understanding of its fundamental structure. Despite recent advances in DNA sequencing, a lack of standardized protocols and common analytical frameworks impedes comparisons among studies, hindering the development of global inferences about microbial life on Earth. Here we present a meta-analysis of microbial community samples collected by hundreds of researchers for the Earth Microbiome Project. Coordinated protocols and new analytical methods, particularly the use of exact sequences instead of clustered operational taxonomic units, enable bacterial and archaeal ribosomal RNA gene sequences to be followed across multiple studies and allow us to explore patterns of diversity at an unprecedented scale. The result is both a reference database giving global context to DNA sequence data and a framework for incorporating data from future studies, fostering increasingly complete characterization of Earth's microbial diversity.
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    A Perspective on Cryptosporidium and Giardia, with an Emphasis on Bovines and Recent Epidemiological Findings
    Abeywardena, H ; Jex, AR ; Gasser, RB ; Rollinson, D ; Stothard, JR (ELSEVIER ACADEMIC PRESS INC, 2015)
    Cryptosporidium and Giardia are two common aetiological agents of infectious enteritis in humans and animals worldwide. These parasitic protists are usually transmitted by the faecal-oral route, following the ingestion of infective stages (oocysts or cysts). An essential component of the control of these parasitic infections, from a public health perspective, is an understanding of the sources and routes of transmission in different geographical regions. Bovines are considered potential sources of infection for humans, because species and genotypes of Cryptosporidium and Giardia infecting humans have also been isolated from cattle in molecular parasitological studies. However, species and genotypes of Cryptosporidium and Giardia of bovids, and the extent of zoonotic transmission in different geographical regions in the world, are still relatively poorly understood. The purpose of this article is to (1) provide a brief background on Cryptosporidium and Giardia, (2) review some key aspects of the molecular epidemiology of cryptosporidiosis and giardiasis in animals, with an emphasis on bovines, (3) summarize research of Cryptosporidium and Giardia from cattle and water buffaloes in parts of Australasia and Sri Lanka, considering public health aspects and (4) provide a perspective on future avenues of study. Recent studies reinforce that bovines harbour Cryptosporidium and Giardia that likely pose a human health risk and highlight the need for future investigations of the biology, population genetics and transmission dynamics of Cryptosporidium and Giardia in cattle, water buffaloes and other ruminants in different geographical regions, the fate and transport of infective stages following their release into the environment, as well as for improved strategies for the control and prevention of cryptosporidiosis and giardiasis, guided by molecular epidemiological studies.
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    The genome and developmental transcriptome of the strongylid nematode Haemonchus contortus
    Schwarz, EM ; Korhonen, PK ; Campbell, BE ; Young, ND ; Jex, AR ; Jabbar, A ; Hall, RS ; Mondal, A ; Howe, AC ; Pell, J ; Hofmann, A ; Boag, PR ; Zhu, X-Q ; Gregory, TR ; Loukas, A ; Williams, BA ; Antoshechkin, I ; Brown, CT ; Sternberg, PW ; Gasser, RB (BMC, 2013)
    BACKGROUND: The barber's pole worm, Haemonchus contortus, is one of the most economically important parasites of small ruminants worldwide. Although this parasite can be controlled using anthelmintic drugs, resistance against most drugs in common use has become a widespread problem. We provide a draft of the genome and the transcriptomes of all key developmental stages of H. contortus to support biological and biotechnological research areas of this and related parasites. RESULTS: The draft genome of H. contortus is 320 Mb in size and encodes 23,610 protein-coding genes. On a fundamental level, we elucidate transcriptional alterations taking place throughout the life cycle, characterize the parasite's gene silencing machinery, and explore molecules involved in development, reproduction, host-parasite interactions, immunity, and disease. The secretome of H. contortus is particularly rich in peptidases linked to blood-feeding activity and interactions with host tissues, and a diverse array of molecules is involved in complex immune responses. On an applied level, we predict drug targets and identify vaccine molecules. CONCLUSIONS: The draft genome and developmental transcriptome of H. contortus provide a major resource to the scientific community for a wide range of genomic, genetic, proteomic, metabolomic, evolutionary, biological, ecological, and epidemiological investigations, and a solid foundation for biotechnological outcomes, including new anthelmintics, vaccines and diagnostic tests. This first draft genome of any strongylid nematode paves the way for a rapid acceleration in our understanding of a wide range of socioeconomically important parasites of one of the largest nematode orders.
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    Next-Generation Molecular-Diagnostic Tools for Gastrointestinal Nematodes of Livestock, with an Emphasis on Small Ruminants: A Turning Point?
    Roeber, F ; Jex, AR ; Gasser, RB ; Rollinson, D (ELSEVIER ACADEMIC PRESS INC, 2013)
    Parasitic nematodes of livestock have major economic impact worldwide. Despite the diseases caused by these nematodes, some advances towards the development of new therapeutic agents and attempts to develop effective vaccines against some of them, there has been limited progress in the development of practical diagnostic methods. The specific and sensitive diagnosis of parasitic nematode infections of livestock underpins effective disease control, which is now particularly important given the problems associated with anthelmintic resistance in parasite populations. Traditional diagnostic methods have major limitations, in terms of sensitivity and specificity. This chapter provides an account of the significance of parasitic nematodes (order Strongylida), reviews conventional diagnostic techniques that are presently used routinely and describes advances in polymerase chain reaction (PCR)-based methods for the specific diagnosis of nematode infections. A particular emphasis is placed on the recent development of a robotic PCR-based platform for high-throughput diagnosis, and its significance and implications for epidemiological investigations and for use in control programmes.
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    Comparative genomics of Cylindrospermopsis raciborskii strains with differential toxicities
    Sinha, R ; Pearson, LA ; Davis, TW ; Muenchhoff, J ; Pratama, R ; Jex, A ; Burford, MA ; Neilan, BA (BMC, 2014-01-29)
    BACKGROUND: Cylindrospermopsis raciborskii is an invasive filamentous freshwater cyanobacterium, some strains of which produce toxins. Sporadic toxicity may be the result of gene deletion events, the horizontal transfer of toxin biosynthesis gene clusters, or other genomic variables, yet the evolutionary drivers for cyanotoxin production remain a mystery. Through examining the genomes of toxic and non-toxic strains of C. raciborskii, we hoped to gain a better understanding of the degree of similarity between these strains of common geographical origin, and what the primary differences between these strains might be. Additionally, we hoped to ascertain why some cyanobacteria possess the cylindrospermopsin biosynthesis (cyr) gene cluster and produce toxin, while others do not. It has been hypothesised that toxicity or lack thereof might confer a selective advantage to cyanobacteria under certain environmental conditions. RESULTS: In order to examine the fundamental differences between toxic and non-toxic C. raciborskii strains, we sequenced the genomes of two closely related isolates, CS-506 (CYN+) and CS-509 (CYN-) sourced from different lakes in tropical Queensland, Australia. These genomes were then compared to a third (reference) genome from C. raciborskii CS-505 (CYN+). Genome sizes were similar across all three strains and their G + C contents were almost identical. At least 2,767 genes were shared among all three strains, including the taxonomically important rpoc1, ssuRNA, lsuRNA, cpcA, cpcB, nifB and nifH, which exhibited 99.8-100% nucleotide identity. Strains CS-506 and CS-509 contained at least 176 and 101 strain-specific (or non-homologous) genes, respectively, most of which were associated with DNA repair and modification, nutrient uptake and transport, or adaptive measures such as osmoregulation. However, the only significant genetic difference observed between the two strains was the presence or absence of the cylindrospermopsin biosynthesis gene cluster. Interestingly, we also identified a cryptic secondary metabolite gene cluster in strain CS-509 (CYN-) and a second cryptic cluster common to CS-509 and the reference strain, CS-505 (CYN+). CONCLUSIONS: Our results confirm that the most important factor contributing to toxicity in C. raciborskii is the presence or absence of the cyr gene cluster. We did not identify any other distally encoded genes or gene clusters that correlate with CYN production. The fact that the additional genomic differences between toxic and non-toxic strains were primarily associated with stress and adaptation genes suggests that CYN production may be linked to these physiological processes.
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    Proteomic Analysis of the Excretory-Secretory Products from Larval Stages of Ascaris suum Reveals High Abundance of Glycosyl Hydrolases
    Wang, T ; Van Steendam, K ; Dhaenens, M ; Vlaminck, J ; Deforce, D ; Jex, AR ; Gasser, RB ; Geldhof, P ; Sripa, B (PUBLIC LIBRARY SCIENCE, 2013-10)
    BACKGROUND: Ascaris lumbricoides and Ascaris suum are socioeconomically important and widespread parasites of humans and pigs, respectively. The excretory-secretory (ES) molecules produced and presented at the parasite-host interface during the different phases of tissue invasion and migration are likely to play critical roles in the induction and development of protective immune and other host responses. METHODOLOGY/PRINCIPAL FINDINGS: The aim of this study was to identify the ES proteins of the different larval stages (L3-egg, L3-lung and L4) by LC-MS/MS. In total, 106 different proteins were identified, 20 in L3-egg, 45 in L3-lung stage and 58 in L4. Although most of the proteins identified were stage-specific, 15 were identified in the ES products of at least two stages. Two proteins, i.e. a 14-3-3-like protein and a serpin-like protein, were present in the ES products from the three different larval stages investigated. Interestingly, a comparison of ES products from L4 with those of L3-egg and L3-lung showed an abundance of metabolic enzymes, particularly glycosyl hydrolases. Further study indicated that most of these glycolytic enzymes were transcriptionally upregulated from L4 onwards, with a peak in the adult stage, particularly in intestinal tissue. This was also confirmed by enzymatic assays, showing the highest glycosidase activity in protein extracts from adult worms gut. CONCLUSIONS/SIGNIFICANCE: The present proteomic analysis provides important information on the host-parasite interaction and the biology of the migratory stages of A. suum. In particular, the high transcriptional upregulation of glycosyl hydrolases from the L4 stage onwards reveals that the degradation of complex carbohydrates forms an essential part of the energy metabolism of this parasite once it establishes in the small intestine.
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    The mitochondrial genome of Protostrongylus rufescens - implications for population and systematic studies
    Jabbar, A ; Mohandas, N ; Jex, AR ; Gasser, RB (BMC, 2013-09-12)
    BACKGROUND: Protostrongylus rufescens is a metastrongyloid nematode of small ruminants, such as sheep and goats, causing protostrongylosis. In spite of its importance, the ecology and epidemiology of this parasite are not entirely understood. In addition, genetic data are scant for P. rufescens and related metastrongyloids. METHODS: The mt genome was amplified from a single adult worm of P. rufescens (from sheep) by long-PCR, sequenced using 454-technology and annotated using bioinformatic tools. Amino acid sequences inferred from individual genes of the mt genomes were concatenated and subjected to phylogenetic analysis using Bayesian inference. RESULTS: The circular mitochondrial genome was 13,619 bp in length and contained two ribosomal RNA, 12 protein-coding and 22 transfer RNA genes, consistent with nematodes of the order Strongylida for which mt genomes have been determined. Phylogenetic analysis of the concatenated amino acid sequence data for the 12 mt proteins showed that P. rufescens was closely related to Aelurostrongylus abstrusus, Angiostrongylus vasorum, Angiostrongylus cantonensis and Angiostrongylus costaricensis. CONCLUSIONS: The mt genome determined herein provides a source of markers for future investigations of P. rufescens. Molecular tools, employing such mt markers, are likely to find applicability in studies of the population biology of this parasite and the systematics of lungworms.
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    Impact of gastrointestinal parasitic nematodes of sheep, and the role of advanced molecular tools for exploring epidemiology and drug resistance - an Australian perspective
    Roeber, F ; Jex, AR ; Gasser, RB (BMC, 2013-05-27)
    Parasitic nematodes (roundworms) of small ruminants and other livestock have major economic impacts worldwide. Despite the impact of the diseases caused by these nematodes and the discovery of new therapeutic agents (anthelmintics), there has been relatively limited progress in the development of practical molecular tools to study the epidemiology of these nematodes. Specific diagnosis underpins parasite control, and the detection and monitoring of anthelmintic resistance in livestock parasites, presently a major concern around the world. The purpose of the present article is to provide a concise account of the biology and knowledge of the epidemiology of the gastrointestinal nematodes (order Strongylida), from an Australian perspective, and to emphasize the importance of utilizing advanced molecular tools for the specific diagnosis of nematode infections for refined investigations of parasite epidemiology and drug resistance detection in combination with conventional methods. It also gives a perspective on the possibility of harnessing genetic, genomic and bioinformatic technologies to better understand parasites and control parasitic diseases.